犬血清和粪便中犬钙卫蛋白定量放射免疫分析方法的建立与分析验证

Development and analytic validation of a radioimmunoassay for the quantification of canine calprotectin in serum and feces from dogs.

作者信息

Heilmann Romy M, Suchodolski Jan S, Steiner Jörg M

机构信息

Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX 77843-4474, USA.

出版信息

Am J Vet Res. 2008 Jul;69(7):845-53. doi: 10.2460/ajvr.69.7.845.

DOI:10.2460/ajvr.69.7.845

PMID:18593232

Abstract

OBJECTIVE

To develop and analytically validate a radioimmunoassay (RIA) for the quantification of canine calprotectin (cCP) in serum and fecal extracts of dogs. Sample Population-Serum samples (n = 50) and fecal samples (30) were obtained from healthy dogs of various breeds and ages.

PROCEDURES

A competitive, liquid-phase, double-antibody RIA was developed and analytically validated by assessing analytic sensitivity, working range, linearity, accuracy, precision, and reproducibility. Reference intervals for serum and fecal cCP concentrations were determined.

RESULTS

Sensitivity and upper limit of the working range were 29 and 12,774 microg/L for serum and 2.9 and 1,277.4 microg/g for fecal extracts, respectively. Observed-to-expected ratios for serial dilutions of 6 serum samples and 6 fecal extracts ranged from 95.3% to 138.2% and from 80.9% to 118.1%, respectively. Observed-to-expected ratios for spiking recovery for 6 serum samples and 6 fecal extracts ranged from 84.6% to 121.5% and from 80.3% to 132.1%, respectively. Coefficients of variation for intra-assay and interassay variability were < 3.9% and < 8.7% for 6 serum samples and < 8.5% and < 12.6% for 6 fecal extracts, respectively. Reference intervals were 92 to 1,121 microg of cCP/L for serum and < 2.9 to 137.5 microg of cCP/g for fecal extracts.

CONCLUSIONS AND CLINICAL RELEVANCE

The RIA described here was analytically sensitive, linear, accurate, precise, and reproducible for the quantification of cCP in serum and fecal extracts. This assay should facilitate research into the clinical use of serum and fecal cCP measurements in dogs with inflammatory bowel disease.

摘要

目的

开发并分析验证一种用于定量犬血清和粪便提取物中犬钙卫蛋白（cCP）的放射免疫分析（RIA）方法。样本群体——从不同品种和年龄的健康犬获取血清样本（n = 50）和粪便样本（30份）。

程序

通过评估分析灵敏度、工作范围、线性、准确性、精密度和可重复性，开发并分析验证了一种竞争性、液相、双抗体RIA方法。确定了血清和粪便cCP浓度的参考区间。

结果

血清的工作范围灵敏度和上限分别为29和12,774μg/L，粪便提取物分别为2.9和1,277.4μg/g。6份血清样本和6份粪便提取物系列稀释的观察值与预期值之比分别为95.3%至138.2%和80.9%至118.1%。6份血清样本和6份粪便提取物加标回收率的观察值与预期值之比分别为84.6%至121.5%和80.3%至132.1%。6份血清样本的批内和批间变异系数分别<3.9%和<8.7%，6份粪便提取物分别<8.5%和<12.6%。血清的参考区间为92至1,121μg cCP/L，粪便提取物为<2.9至137.5μg cCP/g。